Neotropical Entomology ISSN: 1519-566X journal homepage: www.scielo.br/ne

ECOLOGY, BEHAVIOR AND BIONOMICS Survivorship and Egg Produc on of Phytophagous Pentatomids in Laboratory Rearing FAC S1, GS C2, AR P1 1Lab de Bioecologia de Percevejos, Embrapa Soja, Londrina, PR, Brasil 2Univ Filadélfi a de Londrina – UNIFIL, Londrina, PR, Brasil

Keywords Abstract , colony, natural food, reproduc ve performance Survivorship and reproductive performance of the pentatomids Euschistus heros (F.) (EH), (L.) (NV), and Dichelops Correspondence melacanthus (Dallas) (DM) were tested in the laboratory. A mixture A R P, Lab de Bioecologia de of natural foods (pods of green beans, Phaseolus vulgaris, raw shelled Percevejos, Embrapa Soja, CP 231, 86001-970, peanuts, Arachis hypogaea, and fruits of privet, Ligustrum lucidum, Londrina, PR, Brasil; [email protected] and 50 pairs/box (25 x 20 x 20 cm) were used, observed for 30 days, and replicated three times. Thirty days after emergence, mean female Edited by Paulo R Pereira – EMBRAPA survivorship was 91% (EH), 60% (NV), and 30% (DM). More egg Received 14 August 2009 and accepted 27 masses were deposited during 11-20 days after emergence, with January 2010 mean number of 45.1 (EH), 5.3 (NV), and 11.8 (DM). These values were smaller during the irst 10 days (25.5, 2.1, and 4.7) and last 10 days (21-30 days) (39.4, 3.9, and 4.9), respectively. Mean maximum number of eggs/day was 489 (EH) on day 29, 474 (NV) on day 11, and 153 (DM) on day 14. Mean monthly fecundity (egg masses/box) was 985 (EH), 92 (NV), and 193 (DM), and mean number of eggs/ box was 8,480; 5,147, and 2,042.7, respectively.

Introducti on Nezara viridula is known to be reared in the laboratory using raw shelled peanuts and green beans in several Phytophagous pentatomids are important pests causing laboratories worldwide (references in Panizzi et al 2000). economic losses on an array of crops and fruit trees all over However, Brazilian populations of this insect cannot be the world (Panizzi et al 2000). In the Neotropical region, continuously maintained on this diet in laboratory conditions several species are key pests, including the Neotropical (A.R. Panizzi, personal observation). The relative success in brown stink bug, Euschistus heros (F.), and the southern rearing N. viridula is achieved using a combination of whole green stink bug, Nezara viridula (L.), on soybean (Panizzi & soybean plants, dry soybean seeds and raw shelled peanuts Slansky 1985), and Dichelops melacanthus (Dallas) on corn (Corrêa-Ferreira 1985). Euschistus heros is more easily (Ávila & Panizzi 1995, Gomez 1998) and wheat (Chocorosqui reared in the laboratory using green bean pods, dry soybean, & Panizzi 2004, Manfredi-Coimbra et al 2005). peanuts and sunlower seeds (Silva et al 2008), while D. Due to the above mentioned importance of these melacanthus can be maintained on dry soybean seeds and pentatomids, there has been a dramatic increase in the corn seedlings (R. Bianco, personal communication). need of these bugs to perform ield and laboratory trials to This study was conducted to provide information on the select potential insecticides and new varieties. Furthermore, survivorship and reproductive performance (i.e., fecundity) biological control of these bugs using egg parasitoids is an of the three before mentioned pentatomids using a mixture important issue that demands laboratory colonies (Peres & of natural foods, with a selected number of couples/ Corrêa-Ferreira 2004, Silva et al 2008). container for a ixed period of time, in order to establish a Neotrop Entomol 40(1): 35-38 © 2011 Sociedade Entomológica do Brasil 35 Survivorship and Egg Produc on of Phytophagous Pentatomids Silva et al successful routine for rearing these under laboratory difference was observed between the three pentatomid conditions to support egg parasitoid production for the species. The percentage of survivorship of E. heros females implementation of biological control programs. was 91% 30 days after emergence, followed by N. viridula (60%) and D. melacanthus (30%) (Fig 1). Chocorosqui & Panizzi (2008) reported similar survivorship (about 29%) Material and Methods of D. melacanthus fed on mature soybean seed. The mean monthly percentage of female survivorship Insect colony per box was signiicantly higher for E. heros (95.7%), than Adults of E. heros, N. viridula, and D. melacanthus were for N. viridula (78.3%) or D. melacanthus (68.3%) (Table 1). collected at the Embrapa (Empresa Brasileira de Pesquisa Agropecuária) Farm in Londrina Co., northern Paraná State, Brazil (latitude 23º 18’ S), from soybean ields. They were Euschistus heros 100 taken to the laboratory and individualized couples (n = 50) 600 were placed in clear plastic boxes (25 x 20 x 20 cm), and 500 provided with pods of green beans, raw shelled peanuts, and 80 fruits (berries) of privet, Ligustrum lucidum (Oleaceae). Food 400 was replaced every other day. Every two days the egg masses 60 were collected and placed in plastic boxes (11 x 11 x 3 cm), 300 containing moistened ilter paper to prevent desiccation. 40 200 The nymphs were provided with the same food as those 20 used for adults. When nymphs started molting to the ifth 100 Eggs Females instar, they were transferred to plastic boxes (25 x 20 x 20 cm) to complete their development. Boxes were kept in an 0 0 1 3 5 7 9 11131517192123252729 environmental chamber at 26 ± 1°C temperature, 70 ± 10% RH and with a photoperiod of 14:10 h (L:D). Nezara viridula 500 100 Eggs Females Survivorship and reproduc on 450 Survivorship (%) N= 50 400 80 Fifty newly emerged couples of E. heros, N. viridula, and D. 350 melacanthus were selected from the established laboratory 300 60 colony and placed in plastic boxes (25 x 20 x 20 cm) (three 250 boxes for each species), provided with the same food and 200 40 kept at the same environmental condition as the insect 150 colony. A ball (2 cm diameter) of commercial absorbent Mean no. of eggs 100 20 cotton was used as ovipositional substrate for E. heros and 50 D. melacanthus (Silva & Panizzi 2007), and a piece of paper 0 0 towel was provided for N. viridula (Shearer & Jones 1996). 1 3 5 7 9 11131517192123252729 Boxes were daily observed for 30 days and dead females and males were recorded, as well as the number of egg Dichelops melacanthus 180 100 masses and eggs. The mean survivorship (%) of adults and Eggs Females the mean number of egg masses and eggs were calculated 160 80 on a daily basis up to 30 days of adulthood. 140 120 60 Sta s cs 100 80 Data on the total mean number of egg masses and eggs 40 were calculated and submitted to the analysis of variance 60 (ANOVA) and compared using the Tukey test (P < 0.05). 40 20 These analyses were performed using the statistic 20 program SAS 8.2 (SAS Institute 1981, Zar 1984). 0 0 1357911131517192123252729 Days aer emergence Results and Discussion Fig 1 Female survivorship (%) and mean number of eggs Survivorship per day up to 30 days after emergence of three pentatomid species under laboratory conditions (26 ± 1°C, 70 ± 10% Comparing the survivorship of females, a signiicant RH, photophase of 14h). 36 Neotrop Entomol 40(1): 35-38 © 2011 Sociedade Entomológica do Brasil Silva et al Survivorship and Egg Produc on of Phytophagous Pentatomids

Silva et al (2008) observed lower survivorship of E. heros day period of observation and couples should be replaced females (about 60%), in similar conditions, and suggested by younger insects in order to maintain a desirable egg that mortality occurred independently of age. production. Considering the irst 30 days of adulthood, the daily Reproduc ve performance oviposition activity of N. viridula and, to some extent, of D. melacanthus, had occasional pauses or reductions in Euschistus heros produced more egg masses and eggs egg laying activity soon after peaks of activity (Fig 1). This than N. viridula and D. melacanthus in the laboratory behavior was previously reported for N. viridula (Panizzi conditions used in a 30-day period. During the irst 10 & Mourão 1999) and Lygus hesperus (Knight) (Strong et days, E. heros, N. viridula, and D. melacanthus laid a mean al 1970). The egg laying activity of E. heros did not follow number of eggs masses of 25.5, 2.1, and 4.7, respectively. a particular trend. The three pentatomid species deposited more egg masses Comparing monthly fecundity, E. heros deposited during the second 10-day period after emergence (11-20 984.7 egg masses per box, followed by D. melacanthus days), with 45.1 egg masses for E. heros, 5.3 for N. viridula, (193.3) and N. viridula (92). The mean number of eggs and 11.8 for D. melacanthus. These values decreased in the monthly produced per box was 8,480; 5,147, and 2,043 third period (21-30 days) to 39.4, 3.9, and 4.9 egg masses, for E. heros, N. viridula, and D. melacanthus, respectively respectively (Table 2). A similar trend of egg production (Table 1). In similar conditions, Silva et al (2008) reported was observed by Silva et al (2008) when E. heros was a lower egg production by E. heros, but females were reared with different densities of pairs per box. not fed fruits of privet, which is an important host of The highest number of eggs laid/day was 489 eggs several pentatomids species (Panizzi & Grazia 2001). for E. heros on the 29th day, 474 eggs for N. viridula on Nymphs develop well and adults show higher fecundity the 11th day, and 153 eggs for D. melacanthus on the 14th on privet than on other host plants commonly used to day (Fig 1). These results demonstrate that E. heros can rear pentatomids (Panizzi et al 1998, Panizzi & Mourão keep their egg production high over a period of 30 days, 1999, Coombs 2004, Fortes et al 2006). as also observed by Peres & Corrêa-Ferreira (2001) and Euschistus heros females present higher values of Silva et al (2008). On the other hand, N. viridula and D. survivorship and egg production than N. viridula and D. melacanthus decreased their fecundity during the last 10- melacanthus. In soybean ields all over Brazil, E. heros is, in

Table 1 Monthly mean (± SE ) number of female surviving, egg mass size, total number of eggs, and number of eggs laid/female/ box by three pentatomid species under laboratory conditions (26 ± 1°C, RH 70 ± 10%, photophase of 14h).

Monthly (mean ± SE)/box Species Female survivorship (%) Egg mass Eggs Eggs/femalens E. heros 95.7 ± 0.58 a 984.7 ± 8.05 a 8,480.0 ± 67.20 a 187.1 ± 0.49 N. viridula 78.3 ± 2.33 b 92.0 ± 1.01 b 5,147.0 ± 67.44 b 166.0 ± 0.60 D. melacanthus 68.3 ± 4.15 c 193.3 ± 2.19 b 2,042.7 ± 25.00 c 136.2 ± 0.21 Means (n = 3) followed by the same letter in each column do not differ signiicantly using the Tukey test (P > 0.05); nsnon signiicant.

Table 2 Mean (± SE) number of egg masses and eggs laid by pentatomid species in laboratory rearing, up to 30 days after emergence (26 ± 1°C, 70 ± 10% RH, photophase of 14h).

Egg masses (mean ± SE) Eggs (mean ± SE) Time (days) E. heros N. viridula D. melacanthus E. heros N. viridulans D. melacanthus 25.5 ± 7.02 b 2.1 ± 0.80 b 4.7 ± 1.42 b 174.7 ± 48.99 b 143.1 ± 61.00 47.4 ± 14.04 b 0-10 [50] [46] [47] [50] [46] [47] 45.1 ± 2.43 a 5.3 ± 0.49 a 11.8 ± 0.81 a 386.7 ± 21.12 a 282.2 ± 34.56 127.4 ± 7.60 a 11-20 [48] [38] [34] [48] [38] [34] 39.4 ± 2.98 ab 3.9 ± 0.33 ab 4.9 ± 0.79 b 377.5 ± 25.69 a 228.6 ± 24.11 50.2 ± 9.22 b 21-30 [46] [32] [20] [46] [32] [20] Means (n = 3) followed by the same letter in each column do not differ signiicantly using the Tukey test (P > 0.05); nsnon signiicant. Number of females in brackets.

Neotrop Entomol 40(1): 35-38 © 2011 Sociedade Entomológica do Brasil 37 Survivorship and Egg Produc on of Phytophagous Pentatomids Silva et al general, the most abundant stink bug, which suggests that Gomez SA (1998) Controle químico do percevejo Dichelops it is better adapted to different environmental conditions. (Neodichelops) melacanthus (Dallas) (Heteroptera: Pentatomidae) Apparently, it is more resistant to abiotic (temperature, na cultura do milho safrinha. Comunicado Técnico, Embrapa humidity, sun radiation, rain impact, etc.) and biotic Dourados, 44: 1-5. factors (lack of preferred food plants, natural enemies, Manfredi-Coimbra S, Silva JJ, Chocorosqui VR, Panizzi AR (2005) starvation and handling, etc.) than the two other species Danos do percevejo barriga-verde Dichelops melacanthus studied. The fact that N. viridula performed relatively (Dallas) (Heteroptera: Pentatomidae) em trigo. Ciência Rural poorly (and this has been observed in several laboratories 35: 1243-1247. in Brazil) needs to be further investigated, since this Panizzi AR, Grazia J (2001) Stink bugs (Heteroptera, Pentatomidae) species is reared with great success using similar foods and an unique host plant in the Brazilian subtropics. Iheringia in other parts of the world. Ser Zool 90: 21-35. In conclusion, our study demonstrated that the Panizzi AR, McPherson JE, James DG, Javahery M, McPherson RM Neotropical brown stink bug, E. heros is better itted to (2000) Stink bugs (Pentatomidae), p.421-474. In Schaefer CW, be raised in the laboratory than the former two species Panizzi AR (eds) Heteroptera of economic importance. Boca as egg provider for parasitoid rearing. Raton, CRC Press, 828p. Panizzi AR, Mourão APM (1999) Mating, ovipositional rhythm and Acknowledgments fecundity of Nezara viridula (L.) (Heteroptera: Pentatomidae) fed on privet, Ligustrum lucidum Thunb., and on soybean, Glycine max (L.) Merrill fruits. An Soc Entomol Brasil 28: 35-40. We thank D R Sosa-Gómez for critically reading the manuscript. This research was supported by a scholarship Panizzi AR, Mourão APM, Oliveira EDM (1998) Nymph and adult to GSC from the National Council for Scientiic and biology and seasonal abundance of Loxa deducta (Walker) on Technological Development (Conselho Nacional de privet, Ligustrum lucidum. An Soc Entomol Brasil 27: 199-206. Desenvolvimento Cientíico e Tecnológico). This paper Panizzi AR, Slansky Jr F (1985) Review of phytophagous pentatomids was approved for publication by the Editorial Board of (: Pentatomidae) associated with soybean in the Embrapa Soja as manuscript number 16/2009. Americas. Fla Entomol 68: 184-214. Peres WAA, Corrêa-Ferreira BS (2001) Nymphal and adult performance of Euschistus heros (Fabr.) (Hemiptera: References Pentatomidae), as a potential alternative host for egg parasitoids multiplication. Neotrop Entomol 30: 535-540. Ávila CJ, Panizzi AR (1995) Occurrence and damage by Dichelops (Neodichelops) melacanthus (Dallas) (Heteroptera: Pentatomidae) Peres WAA, Corrêa-Ferreira BS (2004) Methodology of mass on corn. An Soc Entomol Brasil 24: 193-194. multiplication of Telenomus podisi Ash. and basalis (Woll.) (: Scelionidae) on eggs of Euschistus heros (Fab.) Chocorosqui VR, Panizzi AR (2004) Impact of cultivation systems (Hemiptera: Pentatomidae). Neotrop Entomol 33: 457-462. on Dichelops melacanthus (Dallas) (Heteroptera: Pentatomidae) populations and damage and its chemical control on wheat. SAS Institute (1981) SAS for linear models. A guide to the ANOVA Neotrop Entomol 33: 487-492. and GLM procedures. SAS Institute, Cary, N.C. Chocorosqui VR, Panizzi AR (2008) Nymphs and adults of Dichelops Shearer PW, Jones VP (1996) Suitability of macadamia nut as a melacanthus (Dallas) (Heteroptera: Pentatomidae) feeding on host plant of Nezara viridula (Hemiptera: Pentatomidae). J Econ cultivated and non-cultivated host plants Neotrop Entomol 37: Entomol 89: 996-1003. 353-360. Silva CC, Laumann RA, Blassioli, MC, Pareja M, Borges M (2008) Coombs M (2004) Broadleaf privet, Ligustrum lucidum Aiton Euschistus heros mass rearing technique for the multiplication (Oleaceae), a late-season host for Nezara viridula (L.), Plautia of Telenomus podisi. Pesq Agropec Bras 43: 575-580. afϔinis Dallas and (Dallas) (Hemiptera: Silva FAC, Panizzi AR (2007) Cotton balls as an oviposition substrate Pentatomidae) in northern New South Wales, Australia. Aust J for laboratory rearing of phytophagous stink bugs (Heteroptera: Entomol 43: 335-339. Pentatomidae). Ann Entomol Soc Am 100: 745-748. Corrêa-Ferreira BS (1985) Criação massal do percevejo verde Strong FE, Sheldahl JA, Hughes PR, Hussein EMK (1970) Reproductive Nezara viridula (L.). Londrina, EMBRAPA-CNPSo, Documentos, biology of Lygus hesperus Knight. Hilgardia 40: 105-147. 11, 16p. Zar JH (1984) Biostatistical analysis, 2nd ed., Prentice-Hall, Fortes P, Magro SR, Panizzi AR, Parra JRP (2006) Development of a dry Englewood Cliffs, N.J. artiicial diet for Nezara viridula (L.) and Euschistus heros (Fabricius) (Heteroptera: Pentatomidae). Neotrop Entomol 35: 567-572.

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